Sensitive Detection of BRAF Hotspot Mutation V600E using E-ice- COLD-PCR Combined with Pyrosequencing

Fei Yu, Kunxian Shu, Dan Pu

2022

Abstract

It is important to sensitively detect BRAF V600E mutation, since it is an important biological marker in several types of cancers, such as melanomas and colorectal cancers. Here, we have combined enhanced improved and complete enrichment co-amplification at lower denaturation temperature-polymerase chain reaction (E-ice-COLD-PCR) with pyrosequencing for the detection of BRAF V600E mutation. A serial of mutation-containing dilutions was determined, and the detect limit of E-ice-COLD-PCR/pyrosequencing and conventional PCR/pyrosequencing assays were 0.1 and 5%, respectively. When BRAF V600E mutation in 10 melanoma patients were further determined, all samples with the V600E mutation detected by conventional PCR/pyrosequencing were also found to be detectable with E-ice-COLD-PCR/pyrosequencing. However, one sample was detected by using E-ice-COLD-PCR/pyrosequencing but not by conventional PCR/pyrosequencing. In summary, E-ice-COLD-PCR/pyrosequencing is sensitive, reliable and cost-effective for detecting BRAF V600E mutation in clinical samples.

Paper Citation

in Harvard Style

Yu F., Shu K. and Pu D. (2022). Sensitive Detection of BRAF Hotspot Mutation V600E using E-ice- COLD-PCR Combined with Pyrosequencing. In Proceedings of the 4th International Conference on Biomedical Engineering and Bioinformatics - Volume 1: ICBEB, ISBN 978-989-758-595-1, pages 535-539. DOI: 10.5220/0011230300003443

in Bibtex Style

@conference{icbeb22,
author={Fei Yu and Kunxian Shu and Dan Pu},
title={Sensitive Detection of BRAF Hotspot Mutation V600E using E-ice- COLD-PCR Combined with Pyrosequencing},
booktitle={Proceedings of the 4th International Conference on Biomedical Engineering and Bioinformatics - Volume 1: ICBEB,},
year={2022},
pages={535-539},
publisher={SciTePress},
organization={INSTICC},
doi={10.5220/0011230300003443},
isbn={978-989-758-595-1},
}

in EndNote Style

TY - CONF

JO - Proceedings of the 4th International Conference on Biomedical Engineering and Bioinformatics - Volume 1: ICBEB,
TI - Sensitive Detection of BRAF Hotspot Mutation V600E using E-ice- COLD-PCR Combined with Pyrosequencing
SN - 978-989-758-595-1
AU - Yu F.
AU - Shu K.
AU - Pu D.
PY - 2022
SP - 535
EP - 539
DO - 10.5220/0011230300003443